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. 2015 Jun 27;71(Pt 7):889–894. doi: 10.1107/S2053230X15008882

Table 2. Overproduction of recombinant AtaPT.

Source organism A. terreus
DNA source cDNA library
Forward primer 5-GCCGGATCCCGGCCCTGGCAGATCC
Reverse primer 5-GCCCTCGAGCACACGTGCGACATTT
Cloning vector pGEX-6P-1
Expression vector pGEX-6P-1
Expression host E. coli BL21 (DE3)
Complete amino-acid sequence of the construct produced MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKKFELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRYGVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVLYMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPKSDLEVLFQGPLGSRPWQILSQALGFPNYDQELWWQNTAETLNRVLEQCDYSVHLQYKYLAFYHKYILPSLGPFRRPGVEPEYISGLSHGGHPLEISVKIDKSKTICRLGLQAIGPLAGTARDPLNSFGDRELLKNLATLLPHVDLRLFDHFNAQVGLDRAQCAVATTKLIKESHNIVCTSLDLKDGEVIPKVYFSTIPKGLVTETPLFDLTFAAIEQMEVYHKDAPLRTALSSLKDFLRPRVPTDASITPPLTGLIGVDCIDPMLSRLKVYLATFRMDLSLIRDYWTLGGLLTDAGTMKGLEMVETLAKTLKLGDEACETLDAERLPFGINYAMKPGTAELAPPQIYFPLLGINDGFIADALVEFFQYMGWEDQANRYKDELKAKFPNVDISQTKNVHRWLGVAYSETKGPSMNIYYDVVAGNVARV

The sequence of the upstream glutathione S-transferase (GST) tag is underlined. PreScission protease can recognize the amino-acid sequence LEVLFQGP and break the peptide bond between Q and G.